We continue to develop our biophysical measurement systems (biological atomic force (Bio-AFM) platforms, Quartz Crystal Microbalance-Dissipation (QCM-D), and optical microscopy and spectroscopy), and to apply these technologies to a number of biomedical and biological investigations in collaboration with NIH intramural and extramural researchers. Major collaborations with notable results in this year include: (A) We have investigated the macromolecular structure and nanomechanical properties of many malaria vaccine candidates via Bio-AFM and related bioanalysis in collaboration with Dr. David Narum (NIAID, NIH), and other co-investigators at the Laboratory of Malaria Immunology and Vaccinology, NIAID, NIH. These protein antigens for malaria vaccine are being produced via recombinant-protein biotechnology, purified, and characterized in a manner suitable for human trials and scale-up productions. We have focused on using AFM and QCM-D to understand the structural properties of these developing vaccines toward enhanced immunological response and eventual eradication of malaria. (B) We have expanded our collaboration on multifunctional nanomedicine probes with Dr. Xiaoyuan Chen (LOMIN, NIBIB), Dr. Ashwin Bhirde (LOMIN, NIBIB), and many co-investigators. New results on several nano-drug and siRNA delivery systems and multi-functional imaging probes have been published and developed further toward cellular imaging and biomedical applications. (C) We have continued our Bio-AFM studies of protein clathrin and clathrin assemblies in collaboration with Dr. Ralph Nossal (NICHD, NIH), Prof. Eileen Lafer (Univ. Texas Health Sciences Center, San Antonio), and coworkers. Clathrin is a key protein for receptor-mediated endocytosis and intracellular trafficking. We continue to used single molecule force spectroscopy(SMFS), AFM imaging and nanomechanical measurements, and QCM-D measurements to characterize clathrin and its assembled structures toward understanding its function from yeast to human. (D) We have advanced a number of other intramural and extramural collaborations involving Bio-AFM and related nanotechnologies for macromolecular and cellular studies.